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Improved protocol for Agrobacterium mediated transformation of tomato and production of transgenic plants containing carotenoid biosynthetic gene CsZCD

TitoloImproved protocol for Agrobacterium mediated transformation of tomato and production of transgenic plants containing carotenoid biosynthetic gene CsZCD
Tipo di pubblicazioneArticolo su Rivista peer-reviewed
Anno di Pubblicazione2007
AutoriQiu, D., Diretto Gianfranco, Tavarza R., and Giuliano Giovanni
RivistaScientia Horticulturae
Volume112
Paginazione172-175
ISSN03044238
Parole chiaveagricultural research, Agrobacterium, Agrobacterium tumefaciens, bacterium, Carotenoid, Crocus, crop production, enzyme, fruit, gene, Lycopersicon esculentum, Rhizobium, transgenic plant
Abstract

Improved protocol for Agrobacterium mediated transformation of tomato (Lycopersicon esculentum) Micro-Tom was developed to use in corporation of the carotenoid biosynthetic genes CsZCD (Crocus zeaxanthin 7,8-cleavage dioxygenase). From these experiment, a transformation methodology using explants from cotyledons cultured for 1 day on the medium with zeatin 2 mg/L, IAA 0.1 mg/L, carefully submerged in the Agrobacterium inoculum for 20 min, then concultured with the agrobacterium for 3 days on the same medium, followed by a transfer to the same medium with 500 mg/L cefotaxin for 3 days and then by a transfer to the same medium with 100 mg/L kanamycin and 500 mg/L carabenillin for 6-8 weeks and resulted in a greater than 20% transformation efficiency in the concentration of Agrobacterium OD600 = 0.2 tested. In this transformation method, no feeder layers were used and the subculture media was minimal. Among the Agrobacterium concentrations of OD600 = 0.2, 0.5 and 1.0, the best transformation efficiency, 20.87%, was obtained by using OD600 = 0.2, which was significantly higher than that of OD600 = 1.0. The presence of the inserted target genes was checked using a rapid and efficient PCR test. The protocol was successfully employed in the production of transgenic Micro-Tom tomato containing the carotenoid biosynthesis CsZCD under constructive promoter. This procedure represents a simple, efficient and general means of transforming tomato. © 2006 Elsevier B.V. All rights reserved.

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URLhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-33847342467&doi=10.1016%2fj.scienta.2006.12.015&partnerID=40&md5=b677e54df27074820c40a6ef843b49db
DOI10.1016/j.scienta.2006.12.015
Citation KeyQiu2007172